W.R. Threlfall

The influence of serum and gonadotropins on in vitro maturation and fertilization of bovine oocytes

The influence of estrous cow serum (ECS) or fetal calf serum (FCS) and their interaction with gonadotropins on in vitro bovine maturation and fertilization was evaluated. The addition of ECS or FCS to the medium significantly increased the percentage of oocyte maturation over that of Hams F-10 medium alone (P<0.05). The addition of follicle stimulating hormone (FSH), human chorionic gonadotropin (hCG) or of FSH plus hCG to the medium provided no advantage in maturation over serum alone. However, FSH plus HCG added to the maturation medium significantly improved the frequency of pronucleus formation in both groups compared with a medium to which no gonadotropins were added (P<0.05). Both ECS and FCS (as medium supplements) promoted oocyte maturation. Although the addition of a combination of FSH plus hCG to the medium did not increase the rate of oocyte maturation, it did contribute to the high incidence of subsequent pronucleus formation.

The effects of estrous cow serum on the in vitro maturation and fertilization of the bovine follicular oocyte

The effect of serum obtained from a cow at the time of standing estrus (serum A), at ovulation (serum B), and at 24 h after ovulation (serum C) on the in vitro maturation and fertilization of bovine oocytes was examined. Of 144 (Group A), 159 (Group B), and 158 (Group C) oocytes, 77 (53.4%), 82 (51.6%) and 82 (51.9%) oocytes were characterized by expansion of cumulus cells, respectively. There was no significant difference in the effect of the three types of cow serum on the cumulus expansion (P < 0.05). Of 461 oocytes, 316 oocytes were cultured with sperm for fertilization, and 145 oocytes were cultured without sperm for evidence of parthenogenetic development. Of 56 (Group A), 56 (Group B), and 62 (Group C) oocytes with expanded cumulus cells, 19 (33.9%), 7 (12.5%), and 11 (17.7%) oocytes were cleaved, respectively, after exposed to the sperm for 24 h. There was a significant difference in the effect of the three types of cow serum on the fertilization rate (P < 0.05). A total of 145 oocytes was cultured in the absence of sperm and no evidence of parthenogenetic division was observed. The effect of the three types of serum obtained from the cow on the maturation of oocytes was not significant, but a significant difference did exist in the fertilization rate of oocytes. Cow serum obtained at the time of standing estrus had a beneficial effect on the fertilization rate of oocytes in vitro.

Histopathologic and gross effects of testicular biopsy in the dog

Eighty-one mixed breed intact male dogs were divided into three groups: Group 1 = left testicle biopsied by Trucut needle method (n=36); Group 2 = left testicle biopsied by Incisional method (n = 39); Group 3 = not biopsied (n = 6). The dogs were castrated 1 to 36 d following biopsy. The biopsy specimens and both testicles were examined for gross and histologic lesions. There was no effect of biopsy on testicle circumference. There was a positive correlation (r(2) = 0.93) between testicular circumference and occurrence of lesions. Incisional biopsies induced more gross lesions (67 vs 33%), and more histologic lesions (135 vs 43). The histologic lesions included hypospermatogenesis, coagulation necrosis, tubular degeneration, interstitial fibrosis and inflammation. In addition, the lesions observed in Incisional biopsy specimens were of greater severity and incidence than Trucut biopsy specimens (87 vs 35%). It was concluded that Incisional biopsy induced an increase in frequency and severity of lesions compared with Trucut biopsy of the canine testicle.

Umbilical blood flow and the effects of premature severance in the neonatal horse

Abstract Thirteen randomly assigned Quarter Horse foals were used in a study to determine the presence of umbilical blood flow immediately post partum and to determine if premature severance of the umbilical cord would alter significantly the hematology of the horse neonate. Packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), hemoglobin concentration (Hgb), red blood cell count (RBC), plasma protein (PP), and heart and respiratory rates were determined every 30 min for the first 6 h of life for both control and treated foals. Control foals had their umbilical cords measured for the presence of umbilical blood flow. Treated foals had their umbilical cords separated after birth and any free flow of blood collected and a volume measure determined. The umbilicus was separated within 10 sec post partum. No blood flow was detected in the control group of foals within 2 min post partum. The blood obtained from the separated umbilicus of the treatment group measured less than 125 cc. PCV, MCV, MCHC, Hgb, RBC, and PP were not significantly different between the two groups (P > 0.05). Except for MCHC and PP, all parameters studied decreased over the first 6 h of life. Heart rate and respiration rates did not significantly differ from each other (P > 0.05) and decreased over the first 6 h of life.

Telemetric monitoring of body temperature in the horse mare

The objective of this study was to develop a radiotelemetric system capable of frequent monitoring of mare body temperature. A radio transmitter was implanted in the flank of each of four mares. Telemetered data were received by a pair of antennae placed at right angles in a 3.3 x 6.6-m stall and stored on a computer hard disk. The data were recorded every 5 minutes except when mares were out of the stall for a 1- to 2- hour exercise period. No effect of environmental temperature, ranging from 5 degrees C to 30 degrees C, on mare body temperature was apparent. The radiotelemetric system used in this study was effective for frequent measurement of mare body temperature.

Equine body temperature and progesterone fluctuations during estrus and near parturition.

Body temperature and serum progesterone concentrations were measured in mares to determine if a change in either could be useful in predicting estrus, ovulation or parturition. There was no significant correlation (P > 0.1) between rectal temperature and the environmental temperature or progesterone concentration. Progesterone concentration did correlate with stage of estrous cycle and the stage of pregnancy. Significant differences (P < 0.05) in temperature were noted at different times throughout the day. No change in temperature occurred that could be utilized to predict estrus, ovulation or parturition. The changes in serum progesterone concentration were only useful in detecting estrus.

The effects of 1,2-propanediol as a cryoprotectant on the freezing of mouse oocytes

Cryopreservation of mammalian eggs has been successfully accomplished using 1,2-propanediol (PG). Effects of holding times of 0 and 30 min at -40 degrees C and storage times of 1 d and 1 mo at -196 degrees C were investigated in combination with various concentrations of PG (1.0, 1.5, and 2.0M) to determine the survival and fertilizability of mouse oocytes rapidly frozen and thawed in straws. A rapid one-step dilution using 0.5 M sucrose solution inside the straws was used following the thawing of oocytes. A significant effect of PG concentration was found between 1.0 M and 1.5 or 2.0 M (P<0.01), but no significance was discovered between 1.5 M and 2.0 M (P>0.05) on subsequent survival and fertilizability of frozen and thawed mouse oocytes. With 2.0 M PG, the best survival rate (58.3%) and fertilizability rate (19.0%) were obtained by holding at -40 degrees C for 30 min and by storage at -196 degrees C for 1 d. Thirty minutes of holding at -40 degrees C reduced oocyte damage during the procedure but not significantly (P>0.05). In addition, there was no significant difference in the various storage periods (P>0.05). This study demonstrated that mammalian oocytes can be cryopreserved in the presence of 1,2-propanediol by utilizing a rapid freezing and thawing procedure.

ABSENCE OF DIURNAL VARIATION IN SERUM PROGESTERONE CONCENTRATIONS IN MARES

Abstract Blood samples were taken at 0800 and 2000 hours each day from 6 horse mares for the duration of one estrous cycle, and from 6 mares for the first 35 days of pregnancy. Progesterone concentration was determined by radioimmunoassay. Data were subjected to repeated measures analyses of variance to compare pregnant vs nonpregnant animals and day and time of day. In addition, within group analyses were performed on data from Days 1 to 18 post ovulation in nonpregnant mares and from Day 1 to 35 post ovulation in pregnant mares. There was no significant effect of time (P=0.1383), and no interaction of time by group (P=0.3225), time by day (P=0.2524), or time by day by group (P=0.5354). Since morning and evening progesterone concentrations did not differ on any day within either group, an average daily progesterone concentration was calculated for each horse on each day. Comparison of daily mean progesterone concentrations (pregnant vs nonpregnant) revealed that pregnant mares did not have higher progesterone concentrations than nonpregnant mares until Days 16, 17 and 18 post ovulation (P There do not appear to be regular diurnal variations in serum progesterone concentrations in the horse mare, although substantial random variation in circulating progesterone can occur throughout the day.

Body temperature fluctuations in the periparturient horse mare

The objective of this study was to determine the pattern of mare deep body temperature fluctuations associated with parturition using biotelemetry. A radio transmitter was implanted in one flank in each of six mares. Telemetered data were received by a pair of antennae placed at right angles in a 3.3 x 6.6-m stall and stored on a computer hard disk. Hourly temperature data were recorded for the period of -168 through 168 hours post partum. A decrease of 0.76 degrees C in body temperature began at 4 hours prior to parturition (P < 0.1) then decreased rapidly between the 3 hours prior to and the time of parturition (Time 0). The lowest mean body temperature recorded was at the time of parturition (36.58 +/- 0.16 degrees C; P < 0.001). A supranormal increase in mean body temperature began one hour post partum, peaked at 38.02 +/- 0.08 degrees C and remained elevated for 48 hours post partum until gradually decreasing to the level of the prepartum mean by 106 hours.

Uterine electromyographic activity in horse mares as measured by radiotelemetry.

The objective of this study was to determine if radiotelemetry could be used to measure myometrial electromyographic (EMG) activity. A radio transmitter with one pair of biopotential leads was implanted in the flank ipsilateral to the pregnant uterine horn at least five weeks prior to the expected date of parturition in two mares. The biopotential leads were implanted in the base of the pregnant uterine horn. Telemetered data were received by a pair of antennae placed at right angles in a 3.3 by 6.6-m stall. Data were recorded on VHS format videocassette tapes continuously for the 24h prior to and following parturition. Simultaneous physiograph recordings were made as a hard copy reference. In addition, 10 mg of prostaglandin F(2 alpha) was administered to two mares in the luteal phase of the estrous cycle. Myometrial EMG during parturition was increased similarly to that of previously published reports that used myometrial electrodes wired directly to a physiograph. Prostaglandin F(2 alpha) also caused an increase in myometrial EMG activity within 8 min of administration. This study demonstrated that radiotelemetry can be used for measuring myometrial EMG activity.